Green fluorescent protein as a marker for expression of a second gene in transgenic plants
Green fluorescent protein as a marker for expression of a second gene in transgenic plants
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DOI：
10.1038/15114
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发表时间：
1999-11-01
影响因子：
46.9
通讯作者：
Stewart, CN
中科院分区：
文献类型：
--
作者：
Harper, BK;Mabon, SA;Stewart, CN
转基因作物的使用引起了人们对转基因转移到非预期宿主及其相关生态后果的关注。此外，转基因表达的田间监测是实际关注的（例如，即将喷洒除草剂的作物中除草剂耐受基因的低表达）。这些潜在问题的解决方案是通过将其与标记基因（如GFP）连接来监测农学上重要的基因的存在和表达。在这里，我们表明，GFP荧光可以指示苏云金芽孢杆菌cry1Ac基因的表达时，共引入烟草和油菜，如昆虫生物测定和蛋白质印迹分析所示。此外，我们进行了两个季节的田间试验，以表征三种不同的GFP基因在转基因烟草中的表现。测试的最佳基因是mGFP5er，一种靶向内质网的诱变GFP基因。我们还证明了在田间合成GFP的宿主植物没有适应性成本。
The use of transgenic crops has generated concerns about transgene movement to unintended hosts and the associated ecological consequences. Moreover, the in-field monitoring of transgene expression is of practical concern (e.g., the underexpression of an herbicide tolerance gene in crop plants that are due to be sprayed with herbicide). A solution to these potential problems is to monitor the presence and expression of an agronomically important gene by linking it to a marker gene, such as GFP. Here we show that GFP fluorescence can indicate expression of the Bacillus thuringiensus cry1Ac gene when co-introduced into tobacco and oilseed rape, as demonstrated by insect bioassays and western blot analysis. Furthermore we conducted two seasons of field experiments to characterize the performance of three different GFP genes in transgenic tobacco. The best gene tested was mGFP5er, a mutagenized GFP gene that is targeted to the endoplasmic reticulum. We also demonstrated that host plants synthesizing GFP in the field suffered no fitness costs.